JPH06501457A - Novel immunosuppressive compounds - Google Patents

Abstract

This invention relates to a novel class of immunosuppressive compounds having an affinity for the FK-506 binding protein (FKBP). Once bound to this protein, the immunosuppressive compounds inhibit the prolyl peptidyl cis-trans isomerase (rotamase) activity of the FKBP and inhibit T cell activation. As such, the compounds of this invention can be used as immunosuppressive drugs to prevent or significantly reduce graft rejection in bone marrow and organ transplantations and for use in the treatment of a wide variety of autoimmune diseases in humans and other mammals.

Description

[Detailed description of the invention] Postoperative graft rejection is a serious complication that affects the success or failure of bone marrow and organ transplants. However, immunosuppressive therapy can significantly reduce graft rejection in organ transplants. can be done.

A variety of diseases are characterized as "autoimmune diseases." These diseases are caused by rejection and reaction. It is similar to graft rejection except that the response is of autologous tissue. immunosuppression Control therapy is also useful in preventing this inappropriate self-rejection.

Syk is one of the widely accepted immunosuppressants for preventing graft rejection. There is Rosporin A (CsA). CsA is a natural fungal metabolite and clinically It has been revealed that Ca1 has strong immunosuppressive activity in organ transplantation. Although it is widely used as a drug, its use (especially at high doses) is associated with nephrotoxicity, It is often associated with side effects such as sexual and other central nervous system disorders.

The following conditions have been treated with cyclosporine A with good results; importance of the autoimmune component in the disease and options similar to tucrosporin A. Effective treatment can be achieved using compounds that act by suppressing T-cell immunosuppression. has been confirmed.

l) Ophthalmology: uveitis, Behcet's disease and Greges' disease.

Note: Nidicrosborin A is currently approved in Japan as a treatment for Behcet's disease. There is. This is the first application of this compound to an autoimmune disease.

2) Dermatology: various autoimmune skin diseases including psoriasis.

. Atopic skin disease.

986). Dry linen.

Ellis, C.N., et al., J., Amer, Med., As5oc, 25 6:3110-3116 (1986). Dryness.

3) Hematology: various diseases including anemia.

Toetterman, T., H., et al., Lancet, 693 (198 4). Pyroblastic wax (PRCA).

Stryckmans, P', A, et al, New Engl, J, Me d, 655-656 (+984). Aplastic anemia.

Gluckman, E. et al., Bone Marrow Transplant 35upp1. 1.2 old (+988). Aplastic anemia.

4) Gastroenterology/Hepatology: Primary liver cirrhosis, autoimmune hepatitis, ulcerative colitis, cancer disease and other gastrointestinal autoimmune diseases.

Wiesner, R.H., et al. Hepatology 7:1025. Abs t, #9. (1987). Primary gallbladder cirrhosis.

Hyams, J.S., et al., Gastroenterology 93:890- 893 (1987).

Autoimmune hepatitis.

, 81lison, M., C. et al., Lancet, 902-903 (19 84). Crohn's disease Brynskov, J. et al., Gastroentero logy92:1330 (1987). Crohn's disease.

Porro, G., B. et al., 1ta1. J, Ga5troentero1 ．． +9:40-41 (+987). Ulcerative colitis.

5) Neurology Amyotrophic lateral sclerosis (ALS, “Lougeli-Tsuku disease”), severe myasthenia and multiple sclerosis.

Appel, S., H., et al., Arch. Neurol., 45:381-386. (+988). amyotrophic lateral sclerosis Tindall, R, S, He, et al, New Engl, J, Med, 3 16:719-724 (1987). Myasthenia gravis. Ann, Neural, 24. No, 1. p, 169. mAbstract P174 (+ 988). Multiple sclerosis.

Dommasch, D. et al. Neurology 385upp1.2.28 -29 (1988). Multiple sclerosis.

6) Nephrotic syndrome, nephrotic syndrome, membranoproliferative glomerulonephritis (MPGN) and related diseases.

Watzon, A., R. et al., Cl1n, Nephrol, 25:273 -274 (1986).

Nephrotic syndrome.

Tejani, A., et al., KidneY rnt, 33 Near 29-734 (1988), Nephrotic syndrome.

Meyer, A., et al., Transplat Proc, 20. 5upp1 ．． 4 (Book rII), 259-261 (1988). nephrosis syndrome.

LaGrue, G. et al., Nephron, 44:382-382 (19 86). M.P.G.N.

7) Osteoarthritis (RA) Harper, J., 1. et al., Lancet 981-982 (1984) , RAoVan Rijthoven, A, W, et al., Ann, Rhe um, Dis, 45 Near 26-731 (1986). R.A.

Dougados, M. et al., Ann, Rheum, Dis. 47:12 7-133 (1988).

R.A.

8) Insulin-dependent diabetes mellitus (rDDM).

Bougneres, P, F, et al., New EngL, J, Med. Town 663-670 (1 Many veterinary diseases are also characterized as autoimmune diseases. . Autoimmune diseases such as those listed above are also observed in mammals [403-410 ．． 491-497 and 498-505 (+986): Cs in canine salivary mammals The mechanism by which A. causes immunosuppression has already been established. In vitro, C sA inhibits the release of lymphokines such as interleukin 2 (IL-2) [Bunjes, D., et al., Eur, J., [mmunol, II:657-6 61 (1981), cloning of helper T cells and cytotoxic T cells. prevent clonal expansion [Larsson, E, J, [mmunol, 124:2828-2833 (1980 )Ko. CsA binds to cyclophilin, a cytosolic protein, and binds to the protein. protein prolyl peptidyl cis-trans isomerase (PPIase) ) Activity 75 (1989)]. PPIase is a peptide bond of prolyl residues. by catalyzing the rotomerization of May be involved in cell activation.

Recently, FK- Another natural product, called 506, is a powerful immune-2 inhibitor and mixed phosphorus Cyclosin inhibits cytotoxic T cell culture responses and inhibits cytotoxic T cell proliferation in vitro. It inhibits more than porin A, but is structurally different from CsA and different from cyclophilin. Binding protein (FKBP) [Harding.

The present invention provides novel proteins with affinity for PK-506 binding protein (FKBP). This invention relates to a group of immunosuppressive compounds. The immunosuppressive compounds of the invention bind to this protein. Then, FKBP prolyl peptidyl cis-trans isomerase guro tamase) activity and inhibit T cell activation. Compounds of the invention prevents or significantly reduces graft rejection in bone marrow and organ transplants immunotherapy and in the treatment of autoimmune diseases in humans and other mammals. It can be used as an epidemic suppressant.

Figure 1 shows preferred compounds of the invention. Each of these preferred compounds The synthesis will be explained in detail in Examples.

Detailed description of the invention The present invention provides novel immunosuppressive compounds of formula I, and pharmacologically acceptable compounds thereof. Regarding these salts.

In the above formula, A represents O, NH, or N-(CI-C4 alkyl), B is hydrogen, CHL-Ar, (CI-C6)-linear or branched alkyl, (C I-C6)-linear or branched alkenyl, (C5-C7)-cycloalkyl , (C5-C7)-cycloalkenyl or (CI-C6)- substituted with Ar alkyl or alkenyl, or In the above formula, L and Q are hydrogen, (CI-C6)-linear or branched atom, respectively. alkyl or (CI-C6)-represents a linear or branched alkenyl, In the above formula, T is Ar, hydrogen, hydroxyl, 0-(CI-C4)-alkyl or from the group consisting of 0-(CI-C4)-alkenyl and carbonyl. Indicates a substituted cyclohexyl having selected substituents at the 3- and 4-positions, where: Ar is hydrogen, halogen, hydroxyl, nitro, CF, (CI-06)-direct Straight-chain or branched alkyl or (CI-C6)-straight-chain or branched alkyl Nyl, 0-(CI-C4)-linear or branched alkyl or ○-(CI-C 4) - linear or branched alkenyl, 0-benzyl, 0-phenyl, amino or 1-naph having 1 to 3 substituents each selected from the group consisting of chill, 2-naphthyl, 2-furyl, 3-furyl, 2-chenyl, 2-pyridyl, selected from the group consisting of 3-pyridyl, 4-pyridyl and phenyl, In the above formula, D is hydrogen or U, and E is oxygen or CH-U. however, When D is hydrogen, E is CH-U; when E is oxygen, D is U , in the above formula, U is hydrogen, O-(CI-C4)-linear or branched alkyl or is O-(CI-C4)-linear or branched alkenyl, (CI-C6)-linear linear or branched alkyl or (CI-C6)-linear or branched alkenyl , (CI-C4)-linear or branched alkyl or (CI-C4)-linear or (C5-C7)-cycloalkyl substituted with branched alkenyl or is (C5-C7)-cycloalkenyl, 2-indolyl, 3-indolyl, [( C1-C4)-alkyl or (CI-C4)-alkenyl)]-Ar or A r (Ar has the same meaning as above), In the above formula, J represents hydrogen, CI or C2 alkyl or benzyl, and K is (C1-C4)-linear or branched alkyl, benzyl or cyclohexy methyl, J and K jointly represent oxygen (0), sulfur (S), S○ or SO A 5- to 7-membered heterocycle which may have 7 substituents may be formed.

The stereochemistry at position 1 (formula I) is (R) or (S), with (S) being preferred.

The compounds of the invention may be used in the form of salts obtained from inorganic or organic acids and bases. can be done. The above acid salts include acetate, ampinate, alginate, Spartate, Benzoate, Henzensulfonate, Bisulfate, Butyrate, Que phosphate, camphorate, camphor sulfonate, cyclopentane propionate, diglu connate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptane acid salt, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, Maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinic acid Salt, oxalate, pamoate, pectate, persulfate , 3-phenylpropionate, picrate, bivalate, propionate, such as succinate, tartrate, thiocyanate, tosylate, and undecanoate. Which can be mentioned. Base salts include ammonium salts, sodium salts and potassium salts. Alkali metal salts such as aluminum salts, alkalis such as calcium salts and magnesium salts Earth metal salts, salts with organic bases such as dicyclohexylamine salts, N-methyl-D - Examples include glucamine and salts with amino acids such as arginine and lysine. Ru. Additionally, these basic nitrogen-containing groups include methyl, ethyl, propyl, and butyl. Lower alkyl halides such as chloride, bromide, iodide, dimethyl sulfate, sulfur Dialkyl sulfates such as diethyl sulfate, dibutyl sulfate and cyamyl sulfate, decyl , lauryl, myristyl, stearyl, chloride, bromide and iodide etc. long-chain halides, aralkyl halogens such as benzyl bromide, phenethyl bromide, etc. It can be quaternized with substances such as compounds. This allows it to dissolve in water or oil. or a dispersed product is obtained.

The compounds of the present invention preferably contain about 750 atomic weight units (a.

m, u, ), most preferably about 500 a, m, υ, or less.

Examples of compounds in which the substituents represented by J and K together form a heterocycle are shown in Table 1. and shown in Table 2. Examples of other preferred compounds of the invention are shown in Tables 3 and 4.

Table 1 2 Bennol phenyl 1 25μM > 5.0μ and co-bennol phenyl 2 1.5 μM > 2.0 μM 4 Allyl phenyl 287aMND5 1 -Naphthyl phenyl 2 0.9μMND6 2-naphthyl phenyl 2 7.0 μM 1.0 μX7 Bennol 2-methyl 1a building 2 0.9 μM HD8 Benzyl 2-methquinophenyl 2 17μ and 75μX9 Ben Zyl 3-methquinophenyl 2 0.3 μ and 1.3 μ and lO benzyl -Methoquinophenyl 2 5.0μM 5.0μH1 Benzyl 3.5-no Methoki/phenyl 2 2.0μM O, 6μM12 beanyl 2,6-, + /fl/phenyl 2 50 μM 25. IJM13 Be, Nor 3. Hey, 5 -To 1j Methoquinophenyl 2 0.1μM to 2μ 14 Beh Le U-ful to Lophenyl 2 4 OμM 10μN 5 Be nor 3-2) lophenyl 2 160μx 75μ and 16 benzyl mu 210 phenyl 2 160μ and yu. OJJM F benzyl 2-bilinol 2 to 130μ > 1ooooμH18 benol 2-Bi9ru N-f+t( F2>500μM 10μM19 tert-7chill 2-7rill l2 0041M > 500IJM Table 1 (continued) 21 Bennol 3-indolyl 2 25μM 20μ and 22 Bennol 2 -f-ophenyl 2 0.8μM 4μH2-enyl phenyl 2 1.5μ M Nil 25 E-3-[Nsu(tophenyl) 2 0.6 μM HD26 E-3-[ ) Lance-(’J-”Nil 2 0.5 μM ND27 Benzyl 2-Nitro Benzyl 1 26 μ to 25 μ 28 Hydrogen Nodokino 2 DoDND 29 tert-butyl metquin 1 600 μM > soo μM 3° Ali Le Metokino 2 190μ and 25μX31 Bennol Metokino 7 2 8 0 years old〉50μH322-/Chlohexylethyl methokino 2　45μM〉 40μ to 33 3-cyclohexylbrobyl methoxy7 2 20μM 12μ X34 1J-Cyclohex/Rubutyl Metquin 2 6pH2-3μH Table 1 ( continuation) propyl 36　E-3-(IJ-men)+n-/)K’i　2　40pM　＞30pH Core E-3-(3,IJ-dime methoxy7 2 10pM ND toquinophenyl )−2= Methyl-prob-2-enyl Core a E-3- (Mu Hydroxy-Haki 7 2 60lM Do D39 E-3- [Nos-(ll-/)Kino 2 70JJM>20JJM42 Bennol 3 -Methoquinphenyl 1 54 E10443 Benzyl 2-bilinol 1 300lMND44'i') Nor 3. Munofluorophenyl 234ND 45 Bennol (E)-2-('I-me)quinophenyl)-214N.

ethinyl 46 Ben 1-hydroxy-1-cyclohexynol 2 14 2μM47 Bennol 2-naphthyl 2 1.5-〇, 3-li48 Bennol 1-naph Chill 2 1lM 2I! M49 (S)-α-methylbennor phenyl 2 0.5p 0.611M5o Bennol 21hydroxy-2-21240, 35$ttetra-dropyranil Table 1 (continued) No, B D nXi:Kd sl (R)-α-methylbennor phenyl 2 13 pM II! M52 Bennol 3-trifluoromethylphenyl 2 1j-M 1.3μM53 Benzyl 3-bennoloxyphenyl 2o・5lM O, 2lM54 Nnor (E)-2-tert-butylethynyl 29μM3I! PI155 Nnor 2-trifluoromethylphenyl 251lMND56 IJ-cyclo Hexylbutyl phenyl 2 0.44 NO57IJ-cyclohexylbutyl 3,11.5-) Rimethquin phenyl 2 0.04lM O,1--58 11-phenylbenzyl phenyl 25IJM N05G’l-phenylbe Njiru 3. IJ, 5-) Rimethquin Phenyl 2 2lMND6o Benzyl □ 3-L) Quinophenyl 2 0j6IAM) JD61 3-phenoquine Bennor 3. Mu, 5-tritquinophenyl 2 0.018lM O,03 5lM62 3-puenoquine benol phenyl 2 0.09lM O, 15 IJM63 4-phenylbutyl 3.11.5-tritquinphenyl 20 ・019lMO・1lM64 Low phenylbutyl phenyl 2 035lM ND65 Bennol 3-(3-probenyloxy)phenyl 2 11. P! N66 <>nor 3-(2-bu○boki/)phenyl 2 0.54 NO 67bennol 1-methylpropyl 2 1B# NO682-phenylbutyl Phenyl 2 1.1lMND69 6-phenylhexyl Phenyl 2 0.5pM ND70 5-phenylpentyl 3. Mu, 5-trimethocinphene Nil 2 0.07. JM Nl) Table 1 (continued) No-B Dn IC-I Xa Mitsu 5-7ri0hexylbenf 3.11.5-trimeth)quinophenyl 2 0.09p 0.08475 Benzyl 3-(1-butoxy)phenyl 2 0.36p N076 Month = Phenylbutyl 3-(2-propoquino)pheni Le 2 0.1p NO7711-(rho-3-dophenyl)butyl 3,11. 5-1 Rimethoquinophenyl 2 0.0161. M O, 06pfu8 Royo Dobennor 3.11.5-1 Rimetokki/Phenyl 2. 41. M ND79 2-(2-naphthyl)ethyl 3. II, 5-) Rimetokki/phenyl 2 0 ．． 224 NDso 2-(+-+)+ru) Echi I & 3. IJ, 5-) Lime eyes Nophenyl 2 0.51. M NDBl 1l-7x Nylbutyl 11-:1 -F7z Nil 2 0. B,! M　0.25p82　Lophenylbutyl　3- Iodophenyl 2 0.13 ta! / 0.2483 3-7. Nirbu 0 Bill 3. Mu, 5-tritankyl phenyl 2 0.11 pM ND84 3-(3-i Nodryl) Propyl 3.mu.5-tritquinophenyl 2 0.017lM O, 054 bowls M! 15 Rho(mumethok11xnyl)butyl 3,11.5 -Trimethquine phenyl 2 0.013 μM O,049p” Moof x 2 1f-2-enyl 3. IJ, 5-) Rimetokki/Phenyl 2 0. JtM ND87 Lauphenylbut-3-enyl 3 mountains 5-trimethquine 7-ninyl 2 　0.51MND! 18 1J-('l-70jlmi/7zJ) Propyl 3. Mu, 5-trinotoki/phenyl 2 0.011p 0.07,1MB9 Mu Phenylbrovir 1-/Chlohexenyl 2 0.78p ND90 IJ- (II-/H)7xnyl)1)-3-enyl 3. IJ, 5-) Rimethokinofe Nil 2 0.011 μM O, 60p91”-phenylpropyl 1-ph Luoro-1-cyclohexyl/ru2 0.54lMND92 11-phenylpro Pill 3-1 Toquinophenyl 2 1.4l MND93 3-[3-(N-nel Mil in drill) Kobrobi 3 3. lJ, 5-) 11 hakin 1 enyl 2 0. 015lM O, D6pA4fi4'I-(3-i>FQk)1+k 3.1 1.5-)'I/)4/7znyl 2 0.016pM 0.05lM95 -Phenylbutyl Bennol 2 0.35lM ND Table 1 (continued) No, B D n Xi E:d 96 11-phenylbutyl 3-biphenyl 2 0JMμM O, mouth 331 M97 8-') x 2 1w square h IJ-tert-buchi 1x:, le 2 0. JtM? JD98 Rhophenylbutyl/Lylohexyl 2 0.08p 0.18 $99”-phenylbutyl cyclohexylmethyl 2 0.1 2$ NDZoo u-1x nylbutyl 3III-methylenenooxyphenyl 2 0.25pM N0101 +1-phenylbutyl rotetrahydropi Ranil 2 0.44pMND103 11-phenylbutyl rho(ll-meth) Toquinphenyloxy-20.7pM ND methyl)-2-7lyl 104 u-7x 21w butchik tert-7 chill 2 0. IJt3ml N O105u-phenylbutyl ethyl 2 1. sz? JD106 3-(N -Benzimidazolyl]propyl 3.11.5-)rimethoxyl phenyl 2 0.11pMNDI07 B-(N-purinyl)propyl 3. u, 5-t I) Tokin 1 enly 2 0.13$ NDlog (S, 5)-2-methyl-3 -Hydroxy-IJ-311115-) Rimeth+17x:k 2 0.254 ND7; Nirbu a building ND means not measured.

Table 2 No, B U n Ki Kd 109 Bennol 31 Mu methylenedioxyphenyl 1 3μH> 15μ 110 Bennol 31u-methyleneoxyphenyl 2 3 μM > 4 μX 111 Bennol Movaquin Phenyl 1 6μ > 30μX112 Ben nor movaquin phenyl 2 4μ to 8μ and 113 benol 2.5- Tsubaki Phenyl 1 10 μMND Uni 4 Bennol 2,11.5-) Lime Toquinophenyl 1 25 μM ND115 Bennol 3. IJ, 5-1 Rimet Quinophenyl 1 450μ and 25μH116 benol munomethylamine Nophenyl 2 20μM > 5μX117 Bennorm-nitrophenyl 2 14 μ to 5 μ 118 Bennol 2-furyl 2 2.5 μM ND 119 Bennol 3-Frill 2 2.5μMND12o Benz' & 3- (F'l 2>604M>8μM121 Bennol 3-pyridyl 2 25 μM ND122 Ben;k Hydrogen 2 KoOOJJM ND Yu23 Be Njiru) Niru 2 Yu 1 “M” - Table 3 No B D JK ICi Kd 125 Bennol Metquin Methyl S-Methyl 400μM > 200μX 126 Bennol Methyl Methyl S-isopropyl 170/JM>2 00μM127 Ethyl Hakino Bennol Hydrogen >1.200μM >30 0 μX u28 ℃ ert-Haxyethyl S-methyl >400 μM >50 07JM Butyl Table 4 No B U S 'KKi Xd 129 Bennol Mumetkin Methyl S-)Til 80 μM > 150 μ to phenyl 1 O benol 4-methquin-methyl S-isopropyl. μ and >20 μ and phenyl Yuko l Bennor 3. +4-methylene-methyl S-methyl 50μMND oxyphenyl 132 Benzyl 3.11-Hlene-hydrogen 5-methyl 60μMND size Nophenyl The immunosuppressive compounds of the present invention can be used in the cytosol of lymphocytes, especially T lymphocytes. sol,) has affinity for the FK-506 binding protein present in

When the immunosuppressive compounds of the present invention bind to FKBP, the prolyl Inhibits peptidyl cis-trans isomerase activity and inhibits FKBP-related It shows the effect of inhibiting lymphocyte activation. One of the FK-506 binding proteins is Hardlng, M.W., et al., Nature 341, 758-760 (19 89) and evaluated the binding affinity of compounds to FKBP. It can be used as a standard for However, the compounds of the present invention It may also show affinity for 6-binding proteins. Prolyl peptidyl Inhibition of cis-trans isomerase is due to binding to PK-506 binding protein It also suggests.

Human FK-506 binding protein is described in Harding, M, W et al., Natu. re 34] Nia 58-760 (1989).

The apparent Kcl value was recorded as 32-l1l-”C)-benzoyl PK-506. It can be used as a reporting ligand for erka, J, J, et al., Nature 341 Near 55-757 (1989 ) using [l HE dihydro-FK-506, Harclin This can be determined by a competitive LH-20 binding assay conducted according to the description of Wear. The binding affinities of several compounds of the present invention to FKBP are shown in Tables 1 to 4. . The data were obtained using the latter method, and [3H]dihydro-PK-5 Competitive ability of unlabeled compounds to bind to FK-506 binding protein of 06 was measured.

Apparent Ki value) is also Harding, M. W., et al., Nature 341 Near 58-760 (1989) or 5iekierka, J., J., et al., Nat. Measured according to the method described in ure 341 near 55-757 (1989). Can be determined. Chymotrypsis releases 4-nitroanilide from the trans form of the substrate In conjunction with tests using the model substrate N-succinyl-Ala- Proline-alanine peptide in Ala-Pro-Phe-p-nitroanilide Cis-trans isomerization of the bond is monitored spectroscopically. Fischer, G. , et al., Nature 337:476-478 (+989). depending on the degree of reaction The inhibitory effect of addition of various concentrations of inhibitor was measured, and the change in the -order rate constant was determined depending on the inhibitor concentration. When analyzed as a function of degree, the apparent Kl value can be evaluated. preferable The degree of enzyme inhibition (Ki) of the compounds is shown in Tables 1 to 4.

The compounds of the present invention are similar in function and use to cyclosporin A and FIG-506. further characterized in in vitro cell biological experiments where it becomes clear that (See Tables 5 and 6).

Table 5 Drugs (Jl crosborin lavaminone FK-506 test and rc so value A 1) Human PBL + < 1μg/ml < 1μg/ml < Tμg/ mlTK3 2) T cell hybrid-7<　Tμg/ml　<　iμg/ml　<　Tμg/ ml+TCR/CD2 3) Apop)-Non 1 μg/ml 1 μg/ml I μg/ml (Apop tosis), inactive in bu07ri, bu0fuku IJ ) CTLL proliferation > 1μg/ml = 0.01Bg/ml >> lμg/ ml+ Engineering L-2 Table 6: Cell test results No, PMA (pM) 0KT3 (μM) LB (μM) JVM (μM) CTLL (μM) 2 ND ND ND ND 3 7.6 4.6 > 10 10 > 8.54 ND ND ND ND NO 5>10>10>10>10>106 ND ND ND ND ND 7>10>10>10>10>108 ND Nl) ND ND N.D. 9>10 6.5>10>io) 1010 ND ND ND ND N.D. ll ND ND ND ND NO 12ND ND ND ND NO 13>10 5.9>10>io>101a ND ND ND ND NO 15ND ND ND ND NO 16) 10 > 10) 10 > 10 > 1017 ND ND ND ND NO 18ND ND ND ND NO 19ND ND ND ND NO 20ND ND ND ND ND Table 6 (continued) 21 ND ND ND ND NO 22:>10>10>10>10>1023>io>10>10 >10 >1024 ND ND ND ND NO 25>10>10>10>10>1025>10>10>10 >10 >1026 >10 6.5 >10 >10 >1027 ND N D ND ND N0 2B ND ND ND ND NO 29ND ND ND ND 30 ND ND ND ND NO 31ND ND ND ND ND 32 ND, ND ND ND ND33 ND ND ND ND ND 34 ND ND ND ND ND 35 ND ND ND ND ND 36 ND ND ND ND ND 37 ND ND ND ND ND 38 ND ND ND ND ND 39 ND ND ND ND ND 40 7.0 1.0 > 10 > 10 > 10 Table 6 (continued) 41 ND ND ND ND ND 42 ND ND ND ND ND 43 ND ND ND ND ND 44 ND ND ND ND ND 45 >10 >10 >10 >10 >1046 >10 >io >10 >10 >1047 >10 >10 >10 >10 >1048 >10 >10 >10 >10 >1049 >10 6.2 >10 >10 > 1050 ND ND ND ND 51 ND ND ND ND ND 52 >10 >10 >10 >10 >1053 >10 El,'O)1 0) 10 B, 054 ND ND ND ND 55 ND, ND ND ND ND56>10:>10>10 6. 5 5.057 4.0 4.5 > 10 8.0 6゜058 ND ND ND ND ND 59 ND ND ND ND ND 60 >10 >10 >10 >10 >10 Table 6 (continued) 61 4.0 B, 0 10 > 10 3.262 6.5 > 10) 10 >10 >1063 6.0 3.1 10 B, 5 3.864 10 6 ．． 0 > 10 > 10 > 1065 > 10) 10 > 10 > 10 > 10 66 >10 >10 >10 >10 >1067 >10 >10 >10 >10 >1068 >10 >10 >10 >10 >1069 6.1 >10 >10 >10 >1070 7.0 4.5 >10 9.0 4 ．． 271 5.0 5.5 7.5 B, 0 3.872 9.0>10> 10>10 5.573 8.0>10 4.5 6.0 7.074 B , 0 9.0 10 10 5.075 B, 0 > 10 9.0 > 10 4 ．． 576 5.0 10 9.0 > 10 6.077 4.5 B, 0 6. 0 7.0 2.17B > 10 > 10 > 10 > 10 > 107910 2.5>jO>10 8.080 3.0 4.0 10 10 6.0 Table 6 (continuation) at 7.0 > 10 > 10 > 10 > 1082 10 > 10) 10 >10 >1083 5.5 5.5 8.5 7.5 5.0B4 4.5 6.0 6.0 > 10 2.085 4.5 4.5 7.0 10 1.5 86 9.0 > 10 > 10 > 10 > 1087 7.0 > 10) 10 >10 >1088 2.2 2.2 2.5 4.5 4.089 8.0 >10 >10 >10 >1090 B, 0 >10) 10 >10 7 ．． 091 >10 >10) 10 >10 >1092 9.0 >10 > 10>10>1093 6.0 7.0 10 B, 7 3.794 5. 0 5.5 > 10 7.0 4095 > 10 > 10 lo > 10 > 1096 >10 >10 >10 >10 >1097 >10 >10 > 10 >10 6.0913 >10 >10 >10 >10 >1099 >10 >10 >10 >10 >10100 >10 >10 >10 > 10 >10101 >10 >10 >10 >10 >10102 >10 >10 10 10 >10 Table 6 (continued) 103 7.0 10 10 > 10 i.

104 4.0 >10 >10 >10 >10105 >10 >10 > 10 >10 >10106 7.0 >10 >io >10 3.0107 6.5 >10 >10 >10 >10108 >10 >10 8.0 >10 >10109 ND ND ND ND NDllOND ND ND ND ND lll ND ND ND ND ND112 ND ND ND ND ND 113 ND ND ND ND ND114 ND ND ND ND 115ND ND ND ND ND 116 ND ND ND ND ND117 ND ND ND ND ND 118ND ND ND ND 119ND ND ND ND 12OND ND ND ND 121ND ND ND ND ND 122ND ND ND ND Table 6 (continued) 123ND ND ND ND ND 124 ND ND ND’～ND ND125 ND ND ND ND ND126ND ND ND ND 127ND NDND ND ND 12BND ND ND ND 129ND ND ND ND ND 130 ND'ND ND ND ND131ND ND ND ND ND 132 ND ND ND No ND All compounds in Table 6 are Toxicity was shown at higher uNa concentrations. It is usually 1 degree higher than 10 μM. Ta.

PMA and 0KT3-mitogens inhibit the proliferation of human peripheral blood lymphocytes (PBCs). used for stimulation. Test compounds were evaluated for their ability to inhibit growth.

Ta.

Inhibition of proliferation of cytotoxic T cells stimulated with CTLL-IL-2! ) Yes hjmura, N. et al. Transplantation 47:356-3 A measurement method similar to that of 59 (+989). The measurements are based on Ficoll-Highback density. Fresh human peripheral blood lymphocytes isolated by high-temperature centrifugation are 0KT3 antibody (anti-CD3) is used for stimulation.

Stimulation was carried out using a radioactive signal with an uninhibited control signal of 48,000-75,000 cpm. Measured by incorporation of thymidine [(“H)TdR] into proliferating cells. IC5 ° values are estimated from the growth inhibition seen at various drug concentrations.

2) Tmm stimulated by anti-T cell receptor (TCR) antibody and anti-CD2 antibody Same measurement method as above except that Tsuclone is used. Stimulus is uninhibited control signal 23. Radioactive thymidine [('H)TdR] as OOOcpm to proliferating cells. Measured by uptake. IC50 value is the growth inhibition seen at various drug concentrations. Evaluate from

3) Shi, Y. et al., Nature 339:625-626 (198 Measurement method according to 9). Measurement uses T cell hybridomas similar to those previously reported. do. T cell hives that mimic effects known to be seen in immature thymocytes. Activation-induced (anti-CD3) cell death in hybridomas (after staining as previously reported) , determined by counting viable cells). Here, cyclo sports The ability of phospho-A and PK-506 to inhibit this cell death is linked to cyclosporine-like and and/or as an indicator for compounds with a PK-506-like mechanism of action. scientific The immunosuppressive drug lavamycin, which is related to, but mechanistically distinct from, this Note that it is inert in the measurements.

4) DuMont, F. et al., J. Immunol, 144+251-25 8 (1990). This measurement method uses CTLL cells that respond to rL-2. Measuring follicular stimulation. Proliferation is measured by (2H)TdR incorporation. S Immunosuppressants that act through a mechanism similar to that of crossporin A and FK-506 are endogenous This IL-2-driven protein works by inhibiting the production of IL-2. It does not show any inhibitory effect on the process. In this measurement method, in order to break down this barrier, using exogenous IL-2. Although chemically related, the mechanisms are clearly different. Note that the immunosuppressant lavamycin is active in this assay. .

These measuring methods can be used to examine the cellular activity of the compounds of the present invention. death Thus, the compounds of the invention are in contrast to the mechanistically distinct immunosuppressive drug lavamycin. However, both cyclosporin A and FK-506 have shown that cellular activities including immunosuppression are It is clear from these results that the results are similar. Furthermore, the cellular activity seen of FEBP binding and inhibition of PPIase (rotamase) activity shown in Table 1. Quantitatively consistent with the activity seen in the case.

Therefore, the compounds of the present invention are useful for preventing organ rejection and treating chronic graft rejection. can be used as an immunosuppressant for the purpose of therapy and the treatment of autoimmune diseases. .

The immunosuppressive compounds of the present invention may be used in patients who have undergone bone marrow transplantation or organ transplantation, or may significantly reduce the patient's immune response, as is the case in various autoimmune diseases, or Can be administered periodically if there is another reason why suppression is desirable. Ru. The compounds of the invention may be used in non-humans for the treatment of various mammalian autoimmune diseases. It can also be administered to other mammals.

Antigen-induced growth and clonal expansion of T cells called ion). This activity is important for the primary prevention of organ transplant rejection, Associated with preservation of transplanted organs during rejection events and inappropriate autoimmune responses It is useful in the treatment of several autoimmune diseases known to be inflammatory. These autoimmune Diseases include uveitis, Behcet's disease, Graves' eye disease, psoriasis, and acute skin disease. Myositis, atopic skin disease, scleroderma, eczema, erythroblastic fistula, aplastic anemia, primary cirrhosis, autoimmune hepatitis, ulcerative colitis, Crohn's disease, amyotrophic lateral sclerosis, Myasthenia gravis, multiple sclerosis, nephrotic syndrome, membranoproliferative glomerulonephritis, deformity These include arthritis and insulin-dependent diabetes. Any of the above autoimmune diseases Treatment is also effective in reducing symptoms and slowing disease progression. in In sulin-dependent diabetes, natural insulin production has completely stopped and exogenous insulin is required. The following treatments are most effective if started before the person becomes completely dependent on phosphorus. is also effective.

For these purposes, the compounds of the invention may be used in conventional non-toxic pharmacologically acceptable compounds. It can be administered orally, parenterally, or by Nebulized, topical, rectal, nasal, buccal, vaginal, or implanted laser Can it be administered via an implanted reservoir? can. As used herein, the term "parenteral" refers to subcutaneous, intravenous, intramuscular , intrasternal, and intracranial injection or infusion techniques.

The pharmaceutical compositions of the invention may be prepared in a sterile form, e.g. as a sterile injectable aqueous or oleaginous suspension. It may also be in the form of an injection preparation. This suspension may be suspended with suitable dispersing or wetting agents. It can be formulated according to methods known per se. The sterile injectable preparation is non-toxic parenterally acceptable, e.g. as a solution in 1,3-butanediol It may also be a sterile injectable solution dissolved or suspended in a diluent or solvent. Available Acceptable vehicles and solvents include water, Ringer's solution, and isotonic saline. Can be mentioned. In addition, sterile, sterile oils are traditionally used as a solvent or suspending medium. has been done. For this purpose, synthetic mono- or di-glycerides and other Mild, non-lucky oils can be used. Fatty acids such as oleic acid and their glycerinase Lido derivatives can be extracted from natural pharmacologically acceptable oils such as olive oil and castor oil, especially Like its polyoxyethylated products, it has applications in the preparation of injectables. these oils Solutions or oil suspensions may contain long chain alcohols such as Ph, He1v and similar alcohols. It may also contain a diluent or a dispersant.

The compounds of the invention may be present, for example, in the form of capsules or tablets, or as an aqueous suspension or It can be administered orally as an aqueous solution. For tablets for oral administration, commonly used Examples of carriers that can be used include lactose and cornstarch. stearic acid Lubricants such as magnesium are also commonly added.

For oral administration in the case of capsule side chambers, useful diluents include lactose and dry coke. Examples include starch. If an aqueous suspension is required for oral use, the active ingredient may be added to milk. Combine with curing and suspending agents. Add some sweeteners and/or colorants if desired. Flavoring and/or coloring agents may also be added.

The compounds of the invention can also be administered in the form of galenic drugs for rectal administration of the drug.

These compositions allow the drug to be solid at room temperature but liquefy at rectal temperature and dissolve in the rectum. prepared by mixing with suitable non-irritating excipients to release the drug Can be done. Such substances include cocoa butter, beeswax, and polyoxyethylene. Examples include lene glycol.

The compounds of the invention are useful for topical administration, particularly for autoimmune diseases of the eye, skin, or lower intestinal tract. It can also be administered locally when the target of treatment is a sensitive area or organ. can. Appropriate topical formulations can easily be prepared for each of these areas.

For ophthalmological use, the compounds of the invention may be used without preservatives such as benzylalkonium chloride. Microparticle suspensions in isotonic, pH-adjusted, sterile saline in the presence or absence of or as a solution in sterile saline, preferably isotonic and pH adjusted. can be done. For ophthalmological use, the compounds of the invention may be formulated in ointments such as petrolatum. You may.

When applied topically to the skin, the compounds of the invention may include, for example, one or more of the following substances: Formulated in the form of a suitable ointment containing the compound suspended or dissolved in a mixture. I can do it. Mineral oil, liquid petrolatum, white petrolatum, propylene glycol, vori Oxyethylene polyoxybrobylene compound, emulsifying wax, water. or, The compounds of the invention may be suspended or dissolved in a mixture of one or more of the following substances: may be formulated as a suitable lotion or cream containing the compound good. Mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester luwax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol Cole, water.

Topical administration to the lower intestinal tract may be in the form of a rectal herbal formulation (see above) or a suitable enema formulation. It can be done with

The above dosage of the active ingredient compound is approximately 0.01 to 1 omg/kg per day. Useful in treating conditions. The active ingredients combined with the carrier material to create a single dose chamber. The amount varies depending on the patient being treated and the method of administration.

That said, the specific dosage for a particular patient will depend on the activity of the compound used, Age, weight, general condition, gender, diet, administration time, excretion rate, drug combination, and It is likely determined by a variety of factors, including the severity of the disease and the severity of the disease being treated.

The compounds of the present invention may be further combined with methylprednisolone acetate to enhance their immunosuppressive effects. It can also be used in conjunction with steroids such as Ron. The steroid can be administered orally or intravenously. , administered rectally, topically, or by inhalation. 0.1-5mg/day kg doses (as methylprednisolone acetate) can be used. 10 An initial loading dose of 0-500 mg can be used. Steroid dosage depends on the clinical situation Depending on the dose, the dose can be reduced from high to low over time. The compound of the invention is , lavamycin, azathioprine, 15- Deoxyspergualine, cyclosporine, PK-506 or a combination thereof It can be administered with other immunosuppressive agents such as Tuclosborin and PK-506 due to contraindications due to co-administration of these immunosuppressants. This has been reported and should be avoided. The dosage of other immunosuppressive drugs depends on the above factors. depends on the immunosuppressive effect of the child and drug combination.

○KT3 is a murine anti-human T lymphocyte CD3 surface antigen monoclonal antibody. , especially for the purpose of relief and recovery from acute allograft rejection in renal transplantation. It can be administered intravenously with other compounds.

The present invention will be explained in more detail with reference to the following examples, but the present invention is not limited to these examples. By way of non-limiting example.

Proton nuclear magnetic resonance ('HN'JR) spectra were obtained by Bruker 300 M+-1 with AC300 device, and also with Pulker AMX500 type equipment. The temperature was measured at 500-tH. The chemical shift for the proton resonance is )J Reported in part vermillion (δ) for e4Si(60,0). analysis High performance liquid chromatography (HPLC) for is performed on one of the Hewlett-Paracard 1050 liquid chromatography devices. I was told. HPLC identification of compounds was performed by Waters Associates Dell. The flow rate was 1.5 ml per minute on a 5 micron, 15 cm column. use The solvent system was A=0.1%H, PO4/H! 0, B=0.1%Hs PO,/ It was CH3CN. 15 minutes linear gradient from 95%A15%B to 100%B It was then run for 1.5 minutes with 100% B and detected at 214 nM.

The compounds described below are illustrated in FIG.

Example 1 (E)-3-[cis-4-(hydroxycyclohexyl)]-]2-methylpro 75 ml of dry Ben-2-enyl N-(phenylglyoxyl)-pibecolei (S)-pipecolic acid (E gbertson, M., and S., J., and Danishefsky, J., Or. 7.3 g (26,1 Add 13.5 ml (0.13 mol) of benzyl alcohol to a slurry of 4 mmol). ) and p-) 5.48 g (28,8 mmo) of luenesulfonic acid monohydrate l) was added. The reaction mixture was refluxed with a Dean-Starck trap. Heated for 2 hours and then cooled to room temperature. This solution is mixed with 400ml of ether. Diluted and stirred at 4°C overnight. The resulting white solid was collected by filtration and Wash with hexane and dry under reduced pressure to remove p-1-rue of benzyl bipecolate. 9.2 g (90%) of the sulfonic acid salt (134) was obtained. 'HNMR (300 MH,,D,0) δ7.63(d), 7°41(S), 7.28(d) , 5.26 (ABq), 4°8 (S), 4.03 (dd), 3.92 (dd) , 3.51-3.39 (01), 3.15-2.93 (s), 2.40 (S) , 2.36-2.24 (m), 1. ．． 98-1.53 (m).

Benzyl bibecolate is a product that completely dissolves a suspension of this salt in ethyl acetate. prepared by conventional methods by treatment with a saturated aqueous solution of sodium bicarbonate until It was done. This aqueous layer was extracted twice with ethyl acetate, the organic layer extract was collected, and MgSO 4, and the solvent was distilled off to give (S)-benzyl bipecolate (133) as a pale Obtained as a yellow oil.

2, (S)-N-(phenylglyoxyl)pipecolic acid (135) 18,0 4.95 g (17.72 mmol) of L-(S)- in ml of methylene chloride Diisopropylethylamide was added to a solution of pipecolic acid tartrate at 0°C. Add 20.4 ml (117,10 mmol) of 7.7 mmol) of chlorotrimethylsilane was added and the resulting solution was diluted with 1 Stir for 5 minutes. Add 17.72 mmol of benzoylformyl chloride to this mixture. was added. The benzoyl formyl chloride was added to the reactor in a separate reaction flask. Dissolved in 18.0 ml of methylene chloride containing a medium amount of dimethylformamide. 2゜66g (17.72mmol) of penzoylformic acid and 2゜ 3 ml (26,37 mmol) of oxalyl chloride at room temperature. It was fresh. The reaction mixture was stirred overnight at 25°C, then 1. ON's Injected into HCl. The aqueous layer was discarded and the organic layer was diluted with a saturated aqueous solution of sodium bicarbonate. Washed twice. Collect the aqueous layer, wash with methylene chloride, and adjust to pH 2.0 with concentrated hydrochloric acid. Acidified and then extracted repeatedly with ether. flash chromatography (Still, W, C, et al, J, Org, Chem, 43:2923 (1978) (1:1 ethyl acetate-hexane containing 1% acetic acid) (S)-N-(phenylglyoxyl) as a mixture of rotamers. 2.3 g of )-pipecolic acid (135) were obtained. 'HNMR (500MHz , CDCl5) δ11.4-11.1 (br S), 8.02 (d), 7 ．． 98 (d), 7.65 (t), 7.58-7.43 (m), 5.4 5 (d), 4゜64 (dd), 4.43 (d), 3.52 (dd), 3.25 (ddd), 3.01 (ddd), 2.42 (d), 2゜24 (d), 1.86-1.78 (m), 1.68-1.38 (m).

3.43 g (21.7 mmol) of cis- and trichloride were added to 45 ml of methylene chloride. Lance-methyl 4-hydroxycyclohexane carboxylate (Noy ce, D, S, Andori, B, Denney, J, Am, Chem , Sac, 74:5912 (1952)) at 0°C. - Addition of 3.0 ml (26,0 mmol) of lutidine followed by 5.5 ml (2 3,8) mmol tert-butyldimethylsilyl trifluoromethane Ruphonate was added. Remove the ice bath and stir the reaction mixture at 25°C for 2 hours. After that, the solution was poured into a saturated aqueous solution of sodium bicarbonate. Distribute into two layers The organic layer was washed with a saturated aqueous solution of steel sulfate, washed with water, and then dried with Mg5Ot. This gave 5.9 g of crude methyl ester(s). 5.72g of this mixture ( A solution of 21.0 mmol) dissolved in 45 ml of anhydrous THF was added to It was treated with 400 mg (10.5 mmol) of Ni hydride.

The reaction mixture was stirred at 25 °C for 0.5 h, then a saturated solution of Rochelle's salt was added. was added slowly to stop the reaction. Dilute the mixture with ether and partition into two layers. Then, the aqueous layer was washed twice with ethyl acetate. Combine the extracts of the organic layer, MgS Dry with O4 and concentrate to obtain 4.9 g of alcohol present as noastereomer. Obtained. Flash chromatography (eluting with 1:5 ethyl acetate-hexane) According to (136) 650mg and (137) 1. Log and mixture of both 2.40g of the product was obtained. Data for (136) 'HNMR (300MH z, CDC1z) δ 3.99-3.92 (m), 3.46 (d), 1. 72-1.58 (m), 1.57-1.36 (m), 0.86 (s), 0.08 (S). Data for (137)・’HNMR(300M Hz, CDCl1) δ 3.47 (dddd), 3.38 (d), 1.8 6-1.67 (m), 1.47-1.16 axis), 1.05-0.77 ( m), 0.72 (s), -0,02 (S).

(465u 1.5.33mmol) of oxa in 5.0ml of methylene chloride Dimethyl sulfoxide (755μ 1. io, 65 mmol) was added. The resulting solution was stirred for 5 minutes, then 650ml of the above alcohol (136) dissolved in 5.0ml methylene chloride g (2.66 mmol) was added. For this reaction, the mixture was heated to -78°C for 45 minutes. After stirring for 2.2 ml. Add (16,0 mmol) of triethylamine The solution was then allowed to warm to room temperature. The reaction was stopped with 1.0NHC1, and the aqueous layer was was extracted with three portions of methylene chloride. The organic extracts were collected and extracted with MgSO4. It was dried and evaporated to dryness to obtain 620 mg of intermediate aldehyde. This is 5.0m (carboethoxyethyridine)triphenylphosphorus in l of methylene chloride 1.22 g (3.36 mmol) of the sample was directly treated. The resulting reaction The mixture was stirred at room temperature overnight and then poured into water. The aqueous layer is divided into two layers. Extracted twice with ethylene chloride. The organic layer was collected, dried with MgS O4, and concentrated. After condensation, 1.55 g of crude product was obtained. Flash chromatography (1.20 300 mg of enoate (138) was eluted with ether-hexane). It was obtained as a solid substance.

5, (E)-3-[cis-(4-tert-butyldimethylsilyloxycyclo hexyl)]-]2-methylprobutu-2-en-1ol (139) 300 mg (0.95 mmol) of Ito (138) was added to 2.0 ml of anhydrous tetra Add 18 mg (0.43 mmol) to a solution in hydrofuran at 25 °C. Add tium aluminum hydride and stir the resulting mixture for 30 minutes did. The reaction was stopped by slowly adding a saturated solution of Rochelle's salt and then Diluted with chill. The two layers were separated and the aqueous layer was extracted with two portions of ethyl acetate.

The combined organic extracts were washed with water and brine, then dried over MgSOa. . The solvent was evaporated and flash chromatography (1:10 ethyl acetate-hexane) 220 mg of allyic alcohol (eluted with lcohol) (139) was obtained. 'HNMR (300MH,, CDC1, )65.34 (d), 3.96 (d), 3°85 (m), 2.26-2. 18 (m), 1.64 (d), 1.61-1.34 (m), 1.82 (s), O, O(S).

68 mg (0.24 mmol) of Allyric Alcohol (139), 42. 3 mg (0, I6 mmol) of acid (+35) and 39.8 mg (0,20 m mol) of 1-(3-dimethylamino-propyl)-3-ethylcarbodiimide Hydrochloride (EDC) was dissolved in 2.0 ml of anhydrous methylene chloride. A catalytic amount of 4-dimethylaminopyridine was added to the solution and the resulting mixture was heated at room temperature. Stir overnight. The reaction mixture was then poured into water to separate into two layers, and the aqueous layer was poured into water. Extracted twice with ethylene chloride. The organic extract layer was collected and dried with MgS O4. , concentrated to give a yellow oil. Flash chromatography (in hexane) (eluting with 15% ethyl acetate) yielded 9.2 mg of ester (140). It was obtained as a mixture of the following compounds. 'HNMR (5001JH2, CDCl δ 8 ．． 01 (d), 7.94 (d), 7.59-7.52 (m), 7.46 -7.39 (m), 7.19 (d), 7.12 (d), 6.82 (d ), 6゜51 (s), 6.38 (S), 5.43 (cl), 4.78 ( ABq), 4.62 (dd), 4°58 (s), 4.41 (d), 3. 51 (dd), 3.23 (ddd), 3.01 (ddd), 2.41 (d), 2.24 (d), 1.91 (S), 1.84-1.76 (m) , 1.65-1.48 (n+), 0.96 (s), 0.18 (S).

Sibrobut-2-enyl N-(phenylglyoxyl)-bipecolei 9.2mg (0.02 mmol) of ester (140) was dissolved in 1.0 ml of acetonitrile. Mix acetonitrile = 48% hydrofluoric acid in a ratio of 95:5 to the solution at 25°C. A solution consisting of a substance is added dropwise and thin layer chromatography shows the disappearance of the raw material. The reaction was carried out with stirring until the end of the reaction. This reaction is started by the addition of a saturated solution of potassium carbonate. It stopped. The reaction mixture was extracted with three portions of ethyl acetate and extracted with MgSO4 After drying, it was concentrated. Flash chromatography (35% acetic acid in hexane) (eluted with ethyl), 6.1 mg (82%) of ester (25) was rotamerized. obtained as a mixture of bodies. 'HNMR (5001JH,, CDC13) δ8. 02 (dd), 7.97 (dd), 7.67-7゜59 (m), 7. 56-7.48 (m), 5.49 (d), 5.44 (d), 4.61 (ABq), 4.44 (S), 4.41 (d), 3.98-3.91 (m ), 3.5 (br d), 3.26 (ddd), 3.11 (ddd), 2.43-2.18 (m), 1.86-1.37 (m), 1.72 (d ), Rf O, 57 (3:1 ethyl acetate-hexane).

Example 2 Freshly prepared (S)-benzylbibecholate (+33) (described in Example 1) 43 mg (0.19 mmol) of was dissolved in 2.0 ml of anhydrous methylene chloride. 44+ng (0.29 mmol) of penzoylformic acid and 5 6 mg (0,29 mmol) of 1-(3-dimethylamino-propyl)-3- Add ethylcarbodiimide hydrochloride (EDC) and mix the resulting mixture. Stir overnight at room temperature. The reaction mixture was poured into water and partitioned into two layers, the aqueous layer being Extracted twice with ethylene chloride. The extracted organic layer was collected and dried with MgS O4. , concentrated to give a yellow oil. Flash chromatography (1:3A) 49 mg (72%) of the keto-amide (3) was recovered by elution with ter-hexane. Obtained as a mixture of transmuters.

'HNltlR (500MH,, CDCl 67.98 (d), 7.91 ( d), 7.58 (t), 7.41-7.30 (m), 5.45 (d) , 5.21 (ABq), 5.06 (ABQ), 4,61 (dd), 4.4 2 (d), 3.48 (dd), 3.19 (ddd), 2.96 (dd d), 2.40 (d), 2.21 (d), 1.83-1.72 (m), 1.61-1.49 (m), 1゜46-1.33 (m), Rf O, 5 5 (1:1 z-thyl-hexane).

Example 3 The above keto-amide (9) was added to 45 mg (0,205 mmol) of (S)-benzi Pibecolate (133) (described in Example 1) and 55 mg (0,306 mmol) l) 3-methoxybenzoylformic acid (Barnish).

T. et al., J. Med. Chem. 24:339 (1981)) Prepared as described in Example 2. Flash chromatography (1:4 (eluted with ether-hexane), 73 mg (93%) of (9) was rotamerized. obtained as a body mixture. 'HNMR (300MH, CDCl,) 67.59 -7.10 (m), 5.42 (d), 5.23 (ABQ), 5.09 (ABq), 4.59 (dd), 4.38 (d), 3.82 (s), 3 ．． 81 (S), 3.48-3.40 (m), 3.20 (ddd), 2.9 8 (ddd), 2.39 (d), 2.21 (d), 1.82-1.70 (m), 1.61-1.22 (m), Rf O, 45 (1:1 ether -hexane).

Example 4 412 mg (1,03 mmol) of (S)benzyl in 40 ml of acetonitrile 198 in a solution of O''C in which bibecholate salt (134) (described in Example 1) was dissolved. μl (1,14 mmol) of diisopropylethylamine, 174 mg (1,2 4 mmol) of α-oxo-2-furan acetic acid, 579 mg ( 1,24 mmol) of benzotriazol-1-yloxytris (dimethylalcohol) Add mino)phosphonium hexafluorophosphate, then 216 μl (1.24 mmol) of diisopropylethylamine was added. the reaction obtained The mixture was stirred at room temperature for 14 hours. It was then evaporated to dryness. 150ml of residue Dissolved in ethyl acetate, 50 ml of 0.5N HC1, 50 ml of saturated NaHCO z, sequentially washed with 50 ml of brine, then dried with MgSO3, then concentrated. did. Flash chromatography (2% ether in methylene chloride) Elution) yielded 163 mg (46%) of the keto-amide (20) as an oil. It was done. This compound (7)'HNMR spectrum (300MHz, CDCl* ) was consistent with the product as a mixture of rotamers. Rf　O,2(methylene chloride 2% ether in Lido). HPLCSRt=+2.83 minutes.

Example 5 145 mg (0.37 mmol) of (S)-benzyl pipecolate salt (134 ) (described in Example 1) in 8.0 ml of methylene chloride. 02 mg (0.57 mmol) of 4-methoxycinnamic acid, 107 mg (0,55 mmol) of 1-(3-dimethylaminopropyl)-3-ethyl Carbodiimide hydrochloride (EDC) and 130 μm (0.74 mmol ) of diisopropylethylamine was added. The resulting solution was stirred at room temperature for 12 hours. Stir and then concentrate under reduced pressure. Flash chromatography (1:1 acetic acid Elution with ethyl-hexane) yielded 91 mg (65%) of the amide (112). Obtained as a colored oil. 'HNIJR (300MHz, CDCIz) δ 7.66 (d), 7.50 (d), 7.35 (m), 6.90 (d), 6.82 (d), 6.63 (d), 5.55 (d), 5.20 (br s ), 4.86 (br s), 4.67 (br d), 4.03 (b r d), 3.83 (S), 3.37 (dt), 2.78 (dt) , 2.33 (br d), 1.74 (m), 1.43 (m). Rf O, 40 (1:1 ether-hexane) Example 6 (ethylenedioxyphenyl)-probu-2-enoylalanine benzyl ester Synthesis of (132) 192 mg (1,0 mmol) of 3,4-methylenedioxycinnamic acid Proline benzyl ester hydrochloride, 121 mg (0,5 mmo 1) Lido and 108 mg (0,5 mmol) of alanine benzyl ester hydrochloride A solution of lido dissolved in 6 ml of acetonitrile was added to 0°35 ml (2,0 mmol). l) of diisopropylethylamine and 443 mg (1°0 mmol) of benzo Triazol-1-yloxytris(dimethylamino)phosphonium hexa Sequential treatment with fluorophosphate. The mixture was stirred for 16 hours and then reduced to Concentrate under pressure. Dissolve the residue in 10 ml of dichloromethane and add 3 times the amount of diethyl injected into ether. This mixture was mixed with water, a 10% solution of potassium bisulfate, water, Washed sequentially with a saturated solution of sodium bicarbonate, water and saturated brine. Organic layer is sulfuric acid Dry over magnesium, filter, and concentrate under reduced pressure. The residue was evaporated into hexafluoride as an eluent. The system uses a stepwise gradient of 30%, 35%, 40%, and 45% ethyl acetate in Purified by silica gel column chromatography. 14 of ester (109) 2 mg was obtained as a colorless oil. Rf=0.4 (40% ethyl acetate in hexane) Chill): HPLC Rt = 12.26: 'HNMR (300MH!) The structure was consistent with that of In addition, 162 mg of ester (132) was converted into a colorless oil. obtained. TLC Rf = 0.22 (40% ethyl acetate in hexane): Rt of HPLC = 11.84 minutes: 'HNMR (300) Jl (,) is the structure It was consistent with the construction.

Example 7 (S)-N-3-(4-methoxyphenyl)-prob-2-enoyl-N-1, 55 g (15.0 mmol) of (S)-N-methylalanine and 9.31 (90 ,0mm.

3.00 g (1 5.8 mmol) of p-1-luenesulfonic acid monohydrate. mixture Items are stored in a Dean-S tark trap. Heated under reflux for 19 hours while removing water. After cooling, the reaction solution was poured into a 200ml evaporator. Upon injection into the solution, a yellow oil precipitated. Decant the solvent and remove the residue. Dissolved in ethyl acetate and concentrated to give a viscous pale yellow oil (141). T.L. C:Rf=0.34 (95:5:0.5 CH2CI2:MeOH:concentrated NH.

OH);'HNMR (300MH!) was consistent with the structure.

96 mg (0.5 mmol) of 4-methquine cinnamic acid and 121 m A suspension of g (0.5 mmol) of (141) in 6 ml dichloromethane was heated with nitrogen. Cooled in an ice water bath under a stream of air. Add this mixture to 0.26 ml (1.5 mmol) ) of diisopropylethylamine and 135 mg (0.53 mmol 1.) of N, N'-bis-(2-oxo-3-oxazolidinyl)phosphinic chloride The mixture was treated and then stirred for 16 hours while slowly warming to room temperature. Triple the mixture of diethyl ether, then water, a 10% solution of potassium bisulfate, water , washed sequentially with a saturated solution of sodium bicarbonate, water and a saturated solution of sodium chloride. did. The organic layer was dried over magnesium sulfate, filtered, and concentrated under reduced pressure. Residue The distillate was prepared using thick layer silica using 40% ethyl acetate in hexane as the eluent. Purified by gel chromatography. 40 mg of ester (129> is yellow) Obtained as an oil. TLC Rf = 0.24 (35% acetic acid in hexane) ethyl): HPLC Rt = i3.34 min, 'HNIilR (300MH , ) was consistent with the structure.

Example 8 (S)-N-methyloxalyl-N-ditylalanine benzyle 848mg ( 2.5 mmol) of (S)-N-Fmoc-N-ethylalanine dichloromethane 436 μl (5.0 mmol) of oxalyl loride followed by a catalytic amount (1 drop) of dimethylformamide. mixture Stirred for 1 hour then concentrated under reduced pressure. Remove the yellow oily residue from the loml ene, followed by 517 mg (0.5 mmol) of benzyl alcohol and 669 m g (5.0 mmol) of silver cyanide. This mixture was heated in an oil bath at 80″C. Heat on top for 20 minutes with vigorous stirring, then cool and pass through a pad of diatomaceous earth. and filtered. The filtrate was concentrated and the residue was eluted with 10% acetone in hexane. The product was purified by silica gel column chromatography. 810 mg of Stell (142) was obtained as a colorless oil. TLC: Rf=0.28 ( 15% acetone in hexane):'HNMR (300 μl, ) is consistent with its structure. did.

Dissolve 0.25 g (0.58 mmol) of (142) in 3 ml acetonitrile. The solution was treated with 3 ml of diethylamine and the mixture was left for 20 hours. Mixed The mixture was concentrated under reduced pressure and the residue was dissolved in 10 ml of acetonitrile and resolventized. was removed. After repeating this operation, dissolve the residue in 4 ml of dichloromethane. Cooled in an ice water bath under a nitrogen stream, and added 111 μl (0.64 mmol) of diiso Propylethylamine treatment followed by 54 μl (0, 64 m mol) of methyl oxalyl chloride. Slowly mix this mixture Stir overnight while warming to room temperature. It was then poured into 3 times the volume of ether. mixture water, a 10% solution of potassium bisulfate, water, a saturated solution of sodium bicarbonate, water , and a saturated solution of sodium chloride. This organic layer is removed using magnesium sulfate. The mixture was dried under vacuum, filtered, and concentrated under reduced pressure. The residue is (1nia) acetone-hexane. It was purified by silica gel chromatography using the San mixture as eluent.

147 mg of ester (+27) were obtained as a colorless product. Rf=O.

36 (35% 7 ml in hexane):/); HPLC Rt = 12.19 min;' HNMR (300 μl, ) was consistent with the structure.

Example 9 (S)-3-cyclopentylpropyl N-(2-methyloxalylthis acid (1 43) is (S)-N-(phenylglyoxyl)pibecolic acid (1 Methyl oxalyl chloride as described in Example 1 for the preparation of 35) Prepared from. That is, 3.16 g (11,32 mmol) of (S)-pipe Colic acid tartrate and 1.19g (12.45mmol) of methyl oxychloride Salyl chloride and 1.25g (51%) of acid (143) as a tan solid obtained as.

'HNMR (300 μl, CDCl5) 65.31 (d), 4.62 (d), 4.49 (brd), 3.61 (br d), 3.90 ( S), 3.88 (S), 3.46 (dt), 2°97 (dt), 2. 40-1.40 (m).

This ester (35) was prepared as 3-cyclopentyl-propyl as described in Example 2. Prepared from ion-1-ol and acid (+43).

Flash chromatography (eluted with 2% ether in methylene chloride) This gave 72 mg (48%) of (35) as a colorless oil. This transformation The 'HNMR spectrum of the compound (300 μl!, CDCl5) shows the rotamers as It was consistent with the production of all Rf = 0.56 (10% ether in methylene chloride ). HPLC Rt = 14.30 minutes.

LeukoPak cells or random normal blood donors (old ■ test) Fresh peripheral blood lymphocytes (PBL, ) are isolated from whole blood with a negative hepatitis test, Histobaku +077 (Sigma chemical Co, St, L ouis, MO) by density centrifugation. Mouse CT LL cytotoxic T-line cells and human Jurkat T-line cells The cells were obtained from ATCC (CTLL-2ATCCT[B214.JUR KAT Clone E6-I ATCCT [B152). To revitalize fresh PBL The human allogeneic B cell line used was EB from normal healthy adult blood donors with two completely different HLA hablotypes. V-transformed lymphocytes. All cell lines are from Gibco Mycotect Test. Mycotect test kit (Gibco Mycotect test kit) Mycoplasma-free, tested for coplasma contamination using standard methods. be. The culture medium contained penicillin (500/ml) and streptomycin (50μ g/ml), L-glutamine 2mM, 2-mercaptoethanol (5XIO ), RPMT16 containing 10% heat-inactivated Fe2 and 10mM HEPES 40 (Gibco, Grand Land, NY).

Compound solution and titration All stock chemical reagents were dissolved in DMSO. Titration of compounds is performed by individual tests. into the culture medium. i.e. 1 μM or 10 gM stock solution Complete RPMI or HB 104 made to final diluted concentration using cumulative 3-fold dilutions from to the medium.

MTT exam The MTT test is performed on proliferating lymphoid or non-lymphoid cell lines. A colorimetric technique for determining the toxicity of test compounds; Various concentrations of the test compound in TetNA by Doria (Biologic Inc.) The viability of cells in the presence or absence of 3-[4,5-dimethyl -Thiazoyl-2-yl 12,5-diphenyl-tetrazolium bromide) was evaluated using 4 hours before the end of the 3-day toxicity test incubation period, the dye MTT Microtiter 20a1 (containing 5mg/ml in PBS pH 7.2). and each well of the cell. At the end of the incubation period, remove most of the medium. Carefully aspirate each well. Then acidified (with 0.04N HC1) Add 100 μl of sopropyl alcohol to dissolve the dye and bring the optical density to 570. Read as the difference between nm and 630 nm (Molecular Devices Thermomax plate reader and Softmax so ftware program, Megulo Park, CA). The results are It was compared with the average OD value of the control group (drug-free medium). and 50% toxic (TCl.

) was calculated.

Mitogenicity test (MitoHenesis As5ays) (“PMA- and 0 KT3”) of human PBLs generated in response to mitogens. Inhibitory effect of test compounds on proliferation (Waithe, W, K, and I (irschhorn, Handbood of Experimental [mmunology, 3rd edition Blackwe [l 5 scientific] Publications, Oxford (1978); MisCA ( (1980)) used various concentrations of test compounds and control drugs (CsA, FK506, PA). gamycin) per well of a 96 large round bottom plate. 5XIO' cells were adjusted to a final volume of 0KT3 ( 10-' final dilution) or PMA (10 ng/n+1) and ionomycin (250ng/ml). 48 hours incubation After hydration (37°C, 15% CO2), cells were treated with 1 μCi of 3H-thymidine. Then, after 24 hours, collect the cells with a Tom Tek cell harvester and LKBβ - Measured with a scintillation counter. Check this result (Cpm) by looking at the culture medium. compared with the control group carried out and the concentration (I C6°) was calculated.

MLR Bio 77 Sei (“LB” and “JVM”) Primary mixed lymphocyte reaction (pri Antigen in mary m1xed IYmphocyte reaction) The proliferation of PBLs activated by rated under or absent. 5X10’ fresh PBLs, 5X10” Mitomycin C treatment of - allogeneic E B V - transformed β-lymphoglastoid cells, L Stimulated with B and JVM (Michelle, B, B, and S, M, Sh iii, 5erected Methods in Ce1lular [ mmunology W, H, Freeman and Co, San Fr Ancisco, CA (1980); Nelson, P.A., et al., Tr. anspIantation 50: 286 (1990)). The exam is 96 A final volume of 200 μl was adjusted per well in large round bottom plates. culture were pulsed on day 6, harvested 24 hours later, and counted as described above.

IL-2 microassay (“CTLL”) test compound is used in the later stages of cytokine utilization. To determine whether to inhibit the T cell activation process, IL-2-dependent 0CTL The proliferative response of the L-20 murine T cell line (ATCC) was evaluated (Gillis, S, et al., J. 1mmunology 120:2027 (1978) ). CsA and FK506 inhibit L-2 production by activated T cells; Lavamycin, on the other hand, interferes with the utilization of Isee2. That is, lavamycin is CT Inhibits IL-2-dependent proliferation of LLs and not CsA and FK506 (Dumont, F, J, et al., [mmunologY +44+25 1 (1990)). 3×10′ CTLLs were transformed into human recombinant [L-2 (Genzyme, rlL-2) at various concentrations in the presence of IU/ml The cells were brought into contact with the compound or control drug for 24 hours. Cells were incubated at lμC 4 hours after drug addition. The ink was pulsed with 3H-thymidine for an additional 20 hours (37°C, 5% C07). It was incubated. Cells were then collected and counted as described above.

Figure IF Figure II Figure IJ Copy and translation of written amendment) Submission (Article 184-8 of the Patent Law) December 29, 1992 −

Claims (31)

[Claims] 1. It has an affinity for FK-506 binding protein and has an affinity for FK-506 binding protein, Immunosuppressant compounds with molecular weights below c. compound). 2. Prolyl peptidyl cis-trans iso of FK-506 binding protein 2. The immunosuppressive compound of claim 1, which is capable of inhibiting merase activity. 3. 2. The immunosuppressive compound of claim 1, having a molecular weight of about 500 amu or less. 4. Compounds with immunosuppressive activity and pharmacologically acceptable compounds represented by the following formula: The salt that is ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ In the above formula, A represents O, NH, or N-(C1-C4 alkyl), In the above formula, B is hydrogen, CHL-Ar, (C1-C6)-linear or branched alkali Kyl, (C1-C6)-linear or branched alkenyl, (C5-C7)-cyc loalkyl, (C5-C7)-cycloalkenyl or substituted with Ar (C1 -C6)-Alkyl or alkenyl, or ▲Mathematical formula, chemical formula, table, etc. ▼ shows, In the above formula, L and Q are hydrogen, (C1-C6)-linear or branched atom, respectively. alkyl or (C1-C6)-linear or branched alkenyl, In the above formula, T is Ar, hydrogen, hydroxyl, O-(C1-C4)-alkyl or from the group consisting of O-(C1-C4)-alkenyl and carbonyl. Indicates a substituted cyclohexyl having selected substituents at the 3- and 4-positions, where: Ar is hydrogen, halogen, hydroxyl, nitro, CF3, (C1-C6)-direct Straight-chain or branched alkyl or (C1-C6)-straight-chain or branched alkeny O-(C1-C4)-linear or branched alkyl or O-(C1-C4 )-linear or branched alkenyl, O-benzyl, O-phenyl, amino and 1-naphthyl having 1 to 3 substituents each selected from the group consisting of 2-naphthyl, 2-furyl, 3-furyl, 2-thienyl, 2-pyridyl, 3 - selected from the group consisting of pyridyl, 4-pyridyl and phenyl, In the above formula, D is hydrogen or U, E is oxygen or CH-U, provided that When D is hydrogen, E is CH-U; when E is oxygen, D is U , in the above formula, U is hydrogen, O-(C1-C4)-linear or branched alkyl or is O-(C1-C4)-linear or branched alkenyl, (C1-C6)-straight chain linear or branched alkyl or (C1-C6)-linear or branched alkenyl , (C1-C4)-linear or branched alkyl or (C1-C4)-linear or (C5-C7)-cycloalkyl substituted with branched alkenyl or is (C5-C7)-cycloalkenyl, 2-indolyl, 3-indolyl, [( C1-C4)-alkyl or (C1-C4)-alkenyl)]-Ar or A indicates r, In the above formula, J represents hydrogen or C1 or C2 alkyl, and K represents (C1-C4 ) - indicates linear or branched alkyl, benzyl or cyclohexylmethyl , J and K jointly have an oxygen (O), sulfur (S), SO or SO2 substituent may form a 5- to 7-membered heterocycle which may be The stereochemistry of is R or S. 5. The immune system according to claim 4, which has an affinity for FK-506 binding protein. Inhibitory compounds. 6. Prolyl peptidyl cis-trans isomerism of FK-506 binding protein 5. The immunosuppressive compound according to claim 4, which is capable of inhibiting enzyme activity. 7. 5. The immunosuppressive compound of claim 4, having a molecular weight of about 750 amu or less. 8. 8. The immunosuppressive compound of claim 7, having a molecular weight of about 500 amu or less. 5. The immunosuppressive compound of claim 4, wherein the carbon at position 9.1 has S stereochemistry. 10. The immunosuppressive compound according to claim 4, wherein J and K are combined and represented by the following formula: , ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ In the above formula, n is 1 or 2. 11. B is benzyl, alkyl, naphthyl, tert-butyl, hydrogen, E-3- Phenyl-2-methyl-prop-2-enyl, E-3-(4-hydroxyphenyl) E-3-(4-hydroxycyclohexyl)-2-methyl-prop-2-enyl, E-3-(4-hydroxycyclohexyl) sil)-2-methyl-prop-2-enyl, cyclohexylethyl, cyclohexyl Cylpropyl, S-sec-phenethyl, cyclohexylbutyl, cyclopentyl Lupropyl, E-3-(4-methoxyphenyl)-2-methyl-prop-2-ethyl E-3-(3,4-dimethoxyphenyl)-2-methyl-prop-2-ethyl Nyl and E-3-[cis-(4-hydroxycyclohexyl)]-2-methyl- selected from the group consisting of prop-2-enyl, and D is phenyl, methoxyphenyl, cyclohexyl, ethyl, methoxy, nitro benzyl, thiophenyl, indolyl, furyl, pyridyl, pyridyl-N-oxy Sid, nitrophenyl, fluorophenyl, trimethoxyphenyl, and dimeth 11. The immunosuppressive compound of claim 10, selected from the group consisting of xyphenyl. 12. The immunosuppressive compound according to claim 4, wherein J and K are combined and represented by the following formula: , ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ In the above formula, n is 1 or 2. 13. U is methoxyphenyl, hydrogen, dimethoxyphenyl, trimethoxyphenyl ru, dimethylaminophenyl, nitrophenyl, furyl, indolyl, pyridyl and methylenedioxyphenyl, according to claim 12. Inhibitory compounds. 14. It is represented by any of the structural formulas shown in Figure 1, has immunosuppressive activity, and A compound that has affinity for FK-506 binding protein. 15. has affinity for FK-506 binding protein and approximately 750 amu An immunosuppressive amount of an immunosuppressive compound having a molecular weight below is physiologically tolerated. A composition (comp) for suppressing an immune response of an individual, which is contained in a vehicle. position). 16. 16. The composition of claim 15, wherein the immune response to be suppressed is an autoimmune response. . 17. The claim is that the immune response to be suppressed is one associated with graft rejection. The composition according to claim 15. 18. Cyclosporine, rapamycin, FK-506, 15-deoxysperg An immunosuppressive substance selected from the group consisting of allin, OKT3 and azathioprine. 16. The composition of claim 15, further comprising: 19. 16. The composition of claim 15, further comprising a steroid. 20. has affinity for FK-506 binding protein and approximately 750 amu The immunosuppressive amount of the compound of claim 4 having a molecular weight of A composition for suppressing an individual's immune response, the composition being contained in a vehicle that can be used to suppress an immune response in an individual. 21. The immunosuppressive compound is one of the compounds listed in Tables 1, 2, 3 or 4. 21. The composition of claim 20. 22. 21. The composition of claim 20, wherein the immunosuppressive compound is represented by the structural formula shown in FIG. . 23. 21. The composition of claim 20, wherein the immune response to be suppressed is an autoimmune response. . 24. The claim is that the immune response to be suppressed is one associated with graft rejection. The composition according to claim 20. 25. has affinity for FK-506 binding protein and approximately 750 amu The immunosuppressive amount of the compound of claim 4 having a molecular weight of during bone marrow transplantation or organ transplantation in an individual, comprising: A composition for preventing or significantly reducing graft rejection. 26. 26. The immunosuppressive compound is represented by the structural formula shown in FIG. Composition of. 27. Cyclosporine, rapamycin, FK-506, 15-deoxysperg An immunosuppressive substance selected from the group consisting of allin, OKT3 and azathioprine. 26. The composition of claim 25, further comprising: 28. 16. The composition of claim 15, further comprising a steroid. 29. has affinity for FK-506 binding protein and approximately 750 amu A physiologically tolerated immunosuppressive amount of the compound of claim 4 having a molecular weight of preventing or significantly reducing an autoimmune response in a mammal. Composition for. 30. 30. The set of claim 29, wherein the immunosuppressive compound is represented by the structural formula shown in FIG. A product. 31. 30. The composition of claim 29, wherein the mammal is a human.